Development of primate cDNA library using cultured cell

我们从以下14种灵长类动物（Pan troglodytes, Pan paniscus, Gorilla Gorilla, Pongo pygmaeus, hyloates syndactylus, hyloates concolor, Cercopithecus mitis, Erythrocebus patas, Presbytis entellus, polykomos, Callithrix jachus, Saguinus oedipus, Tarsius bancanus, Galago senegalensis）中分离的淋巴细胞或成纤维细胞的永活细胞系构建了cDNA文库。为了检测组织非特异性基因的表达，我们检测了cDNA文库中视觉色素基因的表达。在所有库中未检测到表达式。视觉色素是一个基因家族，具有多种光吸收特性，在动物中表现出显著的进化多样性，这可能是由于对特定光环境的适应。一般来说，在一个特定的感光细胞中只表达一种视觉色素。因此，研究视觉色素在动物间的变异及其机制具有重要意义。我们研究了新大陆灵长类动物、啮齿动物、鸟类和鱼类的视觉色素基因的基因组结构和光谱特性。通过分子克隆、Southern杂交、染色体FISH和视觉色素再生等方法，我们发现红-绿视觉色素的三等位基因单位点x染色体模型在血嵌合现象下适用于狨猴。应用同样的方法，我们发现猫头鹰猴在Y染色体上有多个绿色视觉色素基因，其中大部分似乎是无功能的。根据南方杂交的结果，卷尾猴似乎也有两个红绿基因位点。我们克隆了鸽子所有的视觉色素基因，其中一个基因对应于紫外线敏感的视觉色素。测定了小鼠、大鼠和金鱼对紫外色素的吸收光谱。这些结果极大地加深了我们对脊椎动物视觉色素和视觉系统的进化和多样性的理解。少

We have constructed cDNA libraries from immortalized cell lines of lymphocytes or fibroblasts isolated from the following 14 species of primates : Pan troglodytes, Pan paniscus, Gorilla gorilla, Pongo pygmaeus, Hylobates syndactylus, Hylobates concolor, Cercopithecus mitis, Erythrocebus patas, Presbytis entellus, Colobus polykomos, Callithrix jachus, Saguinus oedipus, Tarsius bancanus, and Galago senegalensis. In order to examine tissue-NON-specific gene expression, we tested expression of visual pigment gene in the cDNA libraries. No expression was detected in all libraries. Visual pigments comprise a gene family, which shows a variety of light absorption characteristics, and exhibit remarkable evolutionary diversity among animals possibly due to adaptation to particular light environment. Generally only single type of visual pigment is expressed in a given photoreceptor cell. Therefore, it is of great importance to examine the variation of visual pigments among animals and mechanism … More of cell-type-specific gene expression. We examined genomic organization and spectral properties of visual pigment genes of New World primates, rodents, birds, and fish. By applying molecular cloning, Southern hybridization, chromosome FISH, and visual pigment regeneration to common marmoset, we found that the tri-allelic single-locus X-chromosome model on the red-green visual pigments holds for marmosets under the unique phenomenon, known as blood chimerism. Applying the same methods, we found owl monkeys to have multiple green visual pigment genes on Y chromosome, most of which appeared to be nonfunctional. Capuchin monkeys also appeared to have two red-green gene loci based on the result of Southern hybridization. We cloned all visual pigment genes from pigeon, one of which corresponded to UV-sensitive visual pigment.Absorption spectra of UV pigments were also determined for mouse, rat, and goldfish. These results greatly deepened our understanding of evolution and diversity of vertebrate visual pigments and visual systems. Less